| Citri Reticulatae Pericarpium(CRP),the dried ripe peel of Citrus reticulata Blanco and its cultivars,has been used in foods and drugs due to its effectiveness as regulating qi(energy),normalizing the function of spleen and stomach,resolving phlegm.Liang dynasty Tao Hongjing said,"citrus regulating qi is good,the longer CRP is in storage,the better its efficacy ",For the first time,the phenomenon of CRP should be used,CRP from fresh to old,and the quality gradually improved,is called CRP aging.In the process of aging,CRP is susceptible to mildew and moth,which is closely related to the aging environment.In the previous study of the research group,it was discovered that microflora on the surface of dried tangerine peel could cause changes in the medicinal ingredients.In the previous study of the research group,it was discovered that microflora on the surface of CRP could cause changes in the effective Substance.Therefore,this study investigated the correlations between environmental factors(temperature and humidity),moisture,fungi,and effective Substance components during the aging process of CRP and explained the mechanism of aging.The specific research content is as follows: 1 Drying method of CRPIn December 2016,Cirtus reticu1ata‘Dahongpao' fruit was collected in Xiangfu Town,Jintang County,Sichuan Province,After peeling,dried according to the processing method of the place of origin,dried in the shade,and dried at different temperatures(40°C,60°C),In addition,freeze drying and vacuum drying are also studied.The results showed that drying methods with different moisture content at 40?>dried in the shade>60?>vacuum drying>freeze drying,All of them are in compliance with the moisture regulations under the CRP section of the 2015 edition of the Chinese Pharmacopoeia(?13.0%).Different drying methods have little effect on flavonoids and synephrine,mainly affecting volatile components,and on dry products,volatile oil content freeze drying<dried in the shade<vacuum drying<60?<40?,They are 1.09%,1.11%,1.53%,4.96%,6.33% respectively.Analysis of the composition of volatile components,in addition to the drying method has not been detected,the other drying methods retain the three components in the fresh peel: ?-Humulene,Aromadendrene ?-Elemene ingredients.2 CRP aging process researchFor the first time,the correlation of environmental factors,moisture,fungi,and effective Substance based on the aging process of CRP was studied.The dried samples were aged in a cool,dry,ventilated warehouse and naturally aged for 3 years.Due to time reasons,this study has been conducted for a period of one year,that is,from January 2017 to January 2018.Samples were taken month by month and a total of 13 samples were taken.2.1 Study on the change of environmental factorsUsing a button type temperature-humidity recorder,the environmental factors(temperature and humidity)in the aging process of the dried orange peel are recorded: the temperature in the storage room varies from 3°C to 36.9°C in a year,and the humidity changes from 47% to 89%.In July-October,the temperature is relatively high,and the highest temperature can reach 37°C.In October-December,the humidity is relatively high,and the maximum can reach 89%.The high temperature and high humidity period occurs in September-October,which indicates the change of temperature and humidity in the storage.In line with the climate characteristics of Chengdu: high temperatures in summer and cold in winter.2.2 Water Change ResearchFrom the total moisture and water activity changes,the two changes were more severe.In the summer of July,the moisture content reached a minimum(7.8%),then gradually increased,reaching the maximum(14.0%)in October,and water activity.The minimum was 0.55 in July and 0.71 in November.Therefore,in the storage process should be regularly checked for mildew and insects,especially in September-October,the highest temperature and humidity stage,should be selected when the weather is clear when the sun,in order to prevent mildew.2.3 Study on the Change of Fungal Community StructureFor the first time,Alpha Diversity was used to analyze the richness and diversity of the fungal community during the aging process of CRP.The results showed that the fungal species gradually increased from January to February,and then remained basically unchanged,The species began to decrease gradually in October,and the relative abundance increased gradually from January to April,then decreased,and the abundance increased again from June to September,and then gradually decreased.For the first time,high-throughput sequencing technology was used to analyze the aging process.The main fungal species on the surface of CRP were Ascomycote Microidium spp.,Cladosporium spp.,Candida spp.,Ceramothyrium spp.,and Penicillium spp.,Penicillium spp.,Strelitziana spp.,Aspergillus spp.,Hanseniaspora spp.;Basidiomycota Wallemia spp.,Cyclocybe spp.,This result is basically consistent with the results of the population distribution of the fungal population on the surface of the CRP in the research group,indicating that the fungal group on the surface of the CRP is relatively fixed.During the aging process,the relative abundance of Ascomycota gradually decreased,and the relative abundance of Basidiomycota gradually increased.In October,Aspergillus was detected and its early abundance was low.Analysis of dominant genuses revealed that CRP had more dominant genuses in the months of July to September and February to April.2.4 Study of trait changesObserve and record the appearance color,odor and texture of dried tangerine peel during the aging process,whether there is anti-moisture,mildew,and infestation.At the same time,the surface color values L*(brightness),a*(redness and greenness),and b*(yellowness and blueness)of the CRP were measured using a colorimeter.The results showed that the appearance of the CRP had undergone major changes during its aging.It experienced an orange-red-brown-yellow-brown color change,and the odor gradually faded from the initial richness.The texture was obviously affected by the temperature and humidity in the warehouse.In the high temperature and humidity seasons from August to October,the texture was soft and appeared mild mildew.L*,a*,and b* all showed a gradual decrease,indicating that the appearance of the CRP gradually became darker,and the orange color on the surface gradually shifted toward yellow-brown.This side tendency was consistent with the 2015 edition of the Chinese Pharmacopoeia under a CRP with the appearance of the color matches.2.5 Study on Changes of the Base of effective SubstanceThe content of total flavonoids and polysaccharides was determined by UV spectrophotometry,and the content of flavonoids and synephrine was determined by HPLC.The content of volatile oil was determined by steam distillation method.Static headspace sampling was performed.The dynamic changes of volatile components were determined by GC-MS combined with automated mass spectrometry deconvolution(AMDIS)and Kováts retention index method.The results showed that the contents of total flavonoids,hesperidin,synephrine,and polysaccharides increased.Compared with the month of January,the total flavonoids and polysaccharides increased by 2.6% and 4.8% respectively from the season of aging to December,and the volatile oil yield decreased by 7.7%.The content of nobiletin and tangerine is basically unchanged.From the beginning of aging to the beginning of May,the volatile components of CRP began to change significantly in composition and relative content,resulting in thujone,thymol and other components,while carvacrol,linalylpropionate and other components disappeared.The changes in HPLC fingerprints found that the ratio of compound peak area to its dry product was significantly increased from the beginning of aging to May.And from the beginning of aging to July,a new chemical composition has been produced.Not only does the compound peak area increase and new chemical components are produced.3 Correlation analysis of environmental factors,moisture,fungi and medicinal componentsSpearman correlation was used to analyze the degree of association of temperature,humidity,total moisture,and water activity with fungal community structure.The results showed that the surface of CRP was mainly affected by humidity,water activity,total flavonoids,polysaccharides,of which water activity,total flavonoids significant test P value <0.05,indicating that the higher the surface activity of CRP,the greater the total flavonoid content high,the more abundant the fungus.According to the correlation analysis with dominant genus,it was found that the content of total flavonoids and hesperidin was significantly positively correlated with Wallemia and Aspergillus fungi(P<0.05).There was a significant positive correlation between Wallemia and synephrine(P<0.05),while there was a significant negative correlation between Penicillium,Candida,Aureobasium and total flavonoids,hesperidin,synephrine and polysaccharide content(P< 0.05),indicating that Wallemia,Aspergillus can increase total flavonoids,hesperidin,synephrine content,Penicillium and Candida makes its content reduced.This result can reconfirm the results of the previous research of the research group: The microorganisms on the surface of CRP can cause changes in effective Substance. |
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